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Variability in the resistance of four chicken lines to experimental intravenous infection with Salmonella enteritidis phage type 4.

The capacity of four chicken lines (Y11, L2, B13, PA12) to control Salmonella enteritidis (SE) phage type 4 (PT4) systemic colonization was investigated. Thirteen-week-old chickens were intravenously inoculated with 10(6) SE colony-forming units, and the levels of SE colonization were determined at various time intervals after inoculation in liver, spleen, genital organs, and ceca. The course of SE infection showed a rapid contamination of liver, spleen, and genital organs, whereas the ceca were infected later. A significant (P < 0.001) effect of the chicken line on levels of SE was detected on day 3 postinoculation (PI) in liver and ceca, on day 10 PI in ceca, and on day 15 PI in spleen. Because an early control of systemic Salmonella infection by the Ity/Nramp1 gene has been demonstrated in mice, we aimed to study the early resistance of chickens to SE. As a consequence, we then focused our study on the between- and within-line variabilities of SE levels on day 3 PI. According to the SE levels in liver on day 3 PI, the chicken lines could be classified as susceptible (Y11 and L2) or resistant (PA12 and B13). This early variability was explored in resistant B13 and susceptible L2 lines. Differences between these two lines were confirmed in liver but not in ceca. A large within-line variability was observed in all organs of these two lines. The genetic origin of this variability will have to be determined as a prerequisite to an eventual selection.

Animals↗

Role of Bicaudal-D in patterning the Drosophila egg chamber in mid-oogenesis.

The Bicaudal-D (Bic-D) gene is required early in Drosophila oogenesis for the differentiation of an oocyte from one of a cluster of 16 interconnected germarial cells. To analyze the role of Bic-D later in oogenesis, we have constructed Drosophila lines in which Bic-D expression is under the control of the hsp70 promoter. In these flies, Bic-D activity can be induced early in oogenesis, allowing an oocyte to be made. Then, by shifting females to non-inducing conditions, Bic-D levels are depleted for the remainder of oogenesis. Using this system, we find that Bic-D is indeed required in the later stages of oogenesis. In ovaries from mutant females, oocyte growth is reduced, apparently due to defects in nurse-cell-to-oocyte transport. Smaller oocyte size results in the misalignment of follicle cells and the underlying germ line, leading to ventralization of dorsal follicle cells and to defects in centripetal cell migration. In addition, we show that Bic-D is required for the localization of specific mRNAs at both the anterior and posterior of the oocyte.

Animals↗

Neural circuitry for communication and jamming avoidance in gymnotiform electric fish.

Over the past decade, research on the neural basis of communication and jamming avoidance in gymnotiform electric fish has concentrated on comparative studies of the premotor control of these behaviors, on the sensory processing of communication signals and on their control through the endocrine system, and tackled the question of the degree to which these behaviors share neural elements in the sensory-motor command chain by which they are controlled. From this wealth of investigations, we learned, first, how several segregated premotor pathways controlling a single central pattern generator, the medullary pacemaker nucleus, can provide a large repertoire of behaviorally relevant motor patterns. The results suggest that even small evolutionary modifications in the premotor circuitry can yield extensive changes in the behavioral output. Second, we have gained some insight into the concerted action of the brainstem, the diencephalon and the long-neglected forebrain in sensory processing and premotor control of communication behavior. Finally, these studies shed some light on the behavioral significance of multiple sensory brain maps in the electrosensory lateral line lobe that long have been a mystery. From these latter findings, it is tempting to interpret the information processing in the electrosensory system as a first step in the evolution towards the 'distributed hierarchical' organization commonly realized in sensory systems of higher vertebrates.

Animal Communication↗

Development of free radical products during the greening reaction of caffeic acid esters (or chlorogenic acid) and a primary amino compound.

ESR spectra were measured directly on a marked greening reaction mixture of Et-caffeate and a primary amino compound in alkali solution under aeration. A clear hyperfine structure was commonly detected early in the greening reaction with different amino compounds. Its hyperfine spectrum split into seven peaks was analyzed and found to be due to the oxidized free radical product of the Et-caffeate using an authentic sample system. Another type of hyperfine ESR spectrum was observed later in the reaction, and was altered with different amino compounds. The hyperfine structure for n-butylamine split into 12 lines. The latter type of free radical products were assumed to be a semiquinone type radical compound of the trihydroxy benzacridine derivative, which was identified as the principal structure of the green and yellow pigments formed by this greening reaction system. A formation mechanism of the green pigment and related products involving these free radical products is proposed.

Acridines↗

Infectious human papillomavirus type 31b: purification and infection of an immortalized human keratinocyte cell line.

Human papillomaviruses (HPVs) are aetiological agents of human malignancies, most notably cervical cancers. The life-cycles of HPVs are dependent on epithelial differentiation, and this has impeded many basic studies of HPV biology. The organotypic (raft) culture system supports epithelial differentiation such that infectious virions are synthesized in raft tissues from epithelial cells that replicate extrachromosomal HPV genomes. The CIN-612 9E cell line maintains episomal copies of HPV type 31b (HPV31b), an HPV type associated with cervical cancers. Many previous studies, including our own, have focused on characterizing the later stages of the HPV31b life-cycle in CIN-612 9E raft tissues. In this study, we have used the raft system to generate large numbers of HPV31b viral DNA (vDNA)-containing particles. We found a biologically contained homogenization system to be efficient at virion extraction from raft epithelial tissues. We also determined that vDNA-containing particles could be directly quantified from density-gradient fractions. Using an RT-PCR assay, the presence of newly synthesized, spliced HPV31b transcripts was detected following HPV31b infection of the immortalized HaCaT epithelial cell line. Spliced E6 and E1( wedge )E4 RNAs were detected using a single round of RT-PCR from cells infected with a dose as low as 1.0 vDNA-containing particle per cell. Spliced E1*I,E2 transcripts were found in cells infected with an HPV31b dose as low as 10 vDNA-containing particles per cell. Infectivity was blocked by HPV31 antiserum, but was not affected by DNase I. This work lays a foundation for a detailed analysis of the early events in HPV infection.

3T3 Cells↗

The sensitivity and specificity of the MTS tetrazolium assay for detecting the in vitro cytotoxicity of 20 chemicals using human cell lines.

A number of studies reported that the MTS in vitro cytotoxicity assay is a convenient method for assessing cell viability. The main features found with this assay are its ease of use, accuracy and rapid indication of toxicity. It might well be a useful tool in human health risk assessment if it can be shown that this assay also has an acceptable sensitivity and specificity. This is of interest particularly when exposure to unknown chemical substances requires the rapid detection and evaluation of toxic effects. In this study, the cytotoxicity of 20 chemicals selected from the MEIC priority list was determined with the MTS assay. Since it could be shown that interactions between detection reagents and test chemicals might influence the results of this assay, preliminary experiments were carried out such that artifactual results due to test chemical interference could be excluded from this study. IC50 (50% inhibitory concentration) were established for each test chemical in two human cell lines (F1-73 and HeLa) and later compared with published toxicity data of the same chemicals established with in vitro and in vivo toxicological test systems. Direct comparisons of the data showed a generally lower sensitivity of the MTS assay, which is influenced by biological test organisms, cell type and exposure time. In terms of the specificity of the MTS assay, the results showed a good correlation between data obtained with the MTS assay and published data. The lowest correlation was found when the MTS assay was compared with in vivo studies, however, this finding corresponds well with other published in vitro-in vivo correlations. The highest correlation was found when the MTS assay was compared with test systems using human cell lines or exposure times of 3-24 h. Since the sensitivity of the MTS assay might be increased using different cell types or by extended incubation, this assay is found to provide ideal features of a good measurement system that might also be used for on site toxicological assessments.

Cell Line↗

Spontaneous remission of acute leukemia after the termination of pregnancy.

A 28-year-old woman in the third trimester of her pregnancy was found to have acute myelocytic leukemia. The baby was delivered by Cesarean section and her leukemia underwent spontaneous remission. However, 3 months later, she presented with massive painful leukemic infiltration of the breasts as initial manifestation of relapse, followed by systemic symptoms of leukemia. In vitro culture of the leukemic cells demonstrated characteristics of macrophage cell line. This case illustrates a unique sequence of events: spontaneous remission after the termination of pregnancy, which has profound hormonal alterations, and relapse in a very hormone-sensitive organ, the breast, a few months later when the hormonal milieu was resumed. This suggests hormonal dependence of her leukemic cells and potential for hormonal manipulation in a certain subset of human leukemia.

Adult↗

Lateral asymmetries in infancy: implications for the development of the hemispheres.

Cerebral asymmetry of cognitive processing of stimulus information is commonly viewed as a neocortical phenomenon. However, a number of lines of evidence give innate asymmetry of brainstem motivating systems, which anticipate experience, a key role. Spontaneous asymmetries of gesture and emotion can be observed in infants, who entirely lack language and visuo-constructive skills. Motives for communication in early life may direct subsequent development of complementary cognitive systems in left and right hemispheres. In split-brain monkeys, lateralized motive sets, intentions for manipulation by one hand, can determine which hemisphere will see and learn. Evolutionary antecedents of cerebral asymmetry appear to affect motivation, social signalling and bimanual coordination, with secondary effect in perceptual processing and learning. The hemispheres of adult humans differ in links with neurochemical system that regulate motor initiatives, exploration and attention, and the approach/withdrawal balance in social encounters. Asymmetries in emotional and communicative behaviour in infancy support evidence that an Intrinsic Motive Formation emerging in the embryo human brain stem regulates asymmetries in development and in functioning of the cerebral cortex.

Cerebral Cortex↗

Central projections of the octavolateralis nerves of the clearnose skate, Raja eglanteria.

The central projections of first-order lateral line and octavus nerve afferents of the clearnose skate, Raja eglanteria, were determined by nerve degeneration and horseradish peroxidase techniques. The octavolateralis area of the medulla, which receives these afferents, is organized into dorsal, intermediate, and ventral longitudinal columns of cells and neuropil. Fibers that innervate the electroreceptive sense organs enter the dorsal longitudinal column via the dorsal root of the anterior lateral line nerve and terminate within the dorsal nucleus. Mechanoreceptive fibers from neuromasts of the head and trunk are carried by the ventral root of the anterior lateral line nerve and posterior lateral line nerve, respectively. Both nerves enter the intermediate longitudinal column and terminate throughout the rostrocaudal extent of the intermediate nucleus. Fibers of the ventral root of the anterior lateral line nerve are confined to the medial portion of the intermediate nucleus and posterior lateral line nerve fibers to the lateral portion. In addition, ascending mechanoreceptive fibers from both head and trunk neuromasts project to the vestibulolateral lobe of the cerebellum. Octavus nerve afferents enter the medulla and terminate primarily within the four octaval nuclei that comprise the ventral longitudinal column. Rostrocaudally, these nuclei are the anterior, magnocellular, descending, and posterior octaval nuclei. A few ascending axons continue beyond the anterior octaval nucleus and course to the vestibulolateral lobe of the cerebellum. Some descending axons emanate from the descending octaval nucleus and course to the reticular formation and intermediate nucleus. Therefore, electroreceptive lateral line, mechanoreceptive lateral line, and octavus nerve afferents project ipsilaterally and terminate predominantly within separate medullary nuclei. The significance of octavus nerve projections to the intermediate nucleus and overlap of mechanoreceptive and octavus afferents within the vestibulolateral lobe of the cerebellum cannot be determined until it is known which fibers of the inner ear sense organs project to these areas. Retrograde transport of horseradish peroxidase results in the labeling of large multipolar cells, both ipsilaterally and contralaterally, within a column of gray that lies dorsolateral to the reticular formation. These cells are interpreted as the cell of origin of the efferent components of the anterior and posterior lateral line nerves.

Animals↗

Nephrin expression and three-dimensional morphogenesis of the Xenopus pronephric glomus.

Nephrin (NHPS1) encodes a transmembrane protein of approximately 1,200 amino acids that plays a critical role in podocyte slit-diaphragm formation and the development of functional mammalian glomerular filtration barriers. In humans and mice with congenital defects in the nephrin gene, the glomerular filtration barrier is defective and protein leakage into the kidney filtrate causes a life-threatening proteinuria. This protein also plays an essential role in the formation of the stellate cells of the Drosophila Malpighian tubules. In this report, the sequence and expression of a Xenopus ortholog of nephrin is described using both conventional and novel three-dimensional (3D) visualization methodologies. Xenopus nephrin encodes a protein of 1,238 amino acids and is expressed at high levels in the forming pronephric kidney glomus, the equivalent of the mammalian glomerulus. Expression commences at stage 25 and is specific to the pronephric glomus up until at least tadpole feeding stages. Two-color fluorescent whole-mount in situ analysis of nephrin expression allowed the 3D shape of the glomus to be imaged and contrasted to the pronephric tubules throughout its morphogenesis. Confocal data processing pipelines were established to generate both volumetric and surface models of the developing pronephros, and a Web-based visualization system was used to generate dynamic and manipulable models of the forming nephric organs. This system allows simple on-line morphometric analysis of the developing pronephric components. As in fish embryos, the glomera first form laterally then migrate medially as the pronephros matures. Unlike in the zebrafish, in Xenopus, this migration stops short of complete fusion of the two glomera at the midline, but a nephrin-positive glomeral nexus does form anteriorly and links the two structures from stage 38 onward.

Amino Acid Sequence↗

An experimental polyarthritis associated with systemic antigen excess in immune rabbits.

UNLABELLED: Rabbits were immunized with bovine serum albumin and later challenged intraperitoneally with the same antigen. Polyarthritis, characterized by synovial lining layer hyperplasia and leukocyte infiltration, was conditional on animals having excess free serum antigen in the presence of circulating immune complexes (IC). Nonimmune controls did not develop polyarthritis following antigen challenge. Data are presented that strongly suggest that synovitis was triggered by deposition of IC in the synovia. During the chronic phase of polyarthritis the synovium was clear of antigen but there was marked retention within joint fibrocartilage, suggesting that antigen, immobilized locally, may maintain the chronic synovitis. CLINICAL RELEVANCE: There are several animal models in which experimental arthritis has been induced by antigenic challenge. The resemblance of the lesions to those of human rheumatoid arthritis has provided the rationale for studying the immune events associated with the onset of acute and maintenance of chronic phases of experimental arthritis.

Animals↗

Somatostatin-like immunoreactivity in the brain of an electric fish (Apteronotus leptorhynchus) identified with monoclonal antibodies.

The immunohistochemical localization of somatostatin-like immunoreactive (SSir) cells and fibers in the brain of the gymnotiform teleost (Apteronotus leptorhynchus) was investigated using well-characterized monoclonal antibodies directed against somatostatin-14 and -28. Large populations of SSir neurons occur in the basal forebrain, diencephalon and rhombencephalon and a dense distribution of fibers and terminal fields is found in the ventral, dorsomedial and dorsolateral telencephalon, hypothalamus, centralis posterior thalamus, subtrigeminal nucleus, the motor nucleus of vagus and in the ventrolateral medulla. Immunoreactive neurons in the forebrain are concentrated mainly in the ventral telencephalic areas, the region of the anterior commissure and entopeduncular nucleus. In a fashion similar to the large basal telencephalic cells of other species, the cells of the rostral nucleus entopeduncularis have a significant projection to the dorsal telencephalon. The preoptic region and the peri- and paraventricular hypothalamic nuclei are richly endowed with SSir cells; some of these cells contribute fibres to the pituitary stalk and gland. In the thalamus, only the n. centralis posterior stands out for the density of SSir cells and terminals; these cells appear to project to the prepacemaker nucleus, thus suggesting an SS influence on electrocommunication. In the mesencephalon most SSir cells occur in the optic tectum, torus semicircularis and interpeduncular nucleus. The rhombencephalic SSir cells have a wider distribution (central gray, raphe, sensory nuclei, reticular formation, electrosensory lateral line lobe and surrounding the central canal). The results of this study show the presence of SS in various sensory systems, electromotor system and specific hypothalamic nuclei, suggesting a modulatory role in the processing of sensory information, electrocommunication, endocrine and motor activities.

Animals↗

Moving toward understanding eukaryotic chemotaxis.

The discovery in 1947 of directed cell movement in Dictyostelium discoideum quietly gave a birth to a new line of investigation into the molecular basis of chemotaxis. Some 60 years later, D. discoideum continues to be a key model system for the study of eukaryotic chemotaxis as well as an array of other important biological processes. As one of the most influential scientists, Guenther Gerisch has inspired several generations of researchers with his insightful and rigorous approaches applied to this model system. His studies have greatly contributed to current knowledge of many fundamental processes, such as cell-cell adhesion, phagocytosis, endocytosis, cytokinesis, cell signaling and chemotaxis. In this review, we wish to look back at the journey that has led to our current understanding of chemotaxis of eukaryotic cells.

Animals↗

[Bone stress. A hitherto neglected factor in hip endoprosthesis design].

Growing interest in hip endoprostheses is giving rise to new ideas on form and materials, although it is not precisely known what forces the proximal femur is subject to. To find out more about these forces, we made computerized analyses of the stresses on some different types of prostheses in an artificial standardized femur. The E-module amounts to about 14,000 N/mm. Following measurements in tension-optic trials, we inserted Student gages (EA-13-060LZ-120) at 16 points around the proximal femur; these transmitted the pressure, tensile and torsion stresses to a multipoint measuring instrument (UPM 60) and on-line to a computer. The quality of implantation was monitored by X-ray. With loads varying from 0 to 1.5 kN in 250-N steps we made five readings to reproduce the measurements, without having any material failure. To ensure an absolutely reproducible outcome we used an artificial standardized femur (KNF by Mecron), which without prostheses shows the typical and well-known (Pauwels, Perren) pressure tensions medially and tensile stresses laterally, without crossing the zero line. A distinctly reduced stress profile was seen for pressure and tensile stresses with the ribbed system prosthesis, which might be due to the anchor and collar. Nevertheless, the torsion tensions should be noted. As expected, the shaft systems without anchor and collar show a lesser degree of stiffness. The Mecroblock MR system produces preponderantly pressure stresses, while the CLS (Spotorno) prosthesis sets up more torsion tensions in different directions.(ABSTRACT TRUNCATED AT 250 WORDS)

Biomechanical Phenomena↗

Usefulness of three-dimensional non-fluoroscopic mapping in the ablation of typical atrial flutter.

BACKGROUND: Catheter ablation of the cavo-tricuspid isthmus is rapidly becoming the first line of treatment in the management of atrial flutter. The standard procedure is usually performed under fluoroscopy guidance and relays on multisite endocardial recordings to assess the completeness of the isthmus conduction block. Despite the high rate of success there is, at follow-up, a considerable number of recurrences which could reflect the limitations of conventional assessment of conduction block across the isthmus. This new non-fluoroscopic mapping system allowing high density mapping along the entire length of the ablation line, could provide a more accurate way of verifying complete conduction block. The aim of the present study was to describe our overall results and long-term follow-up using a three-dimensional mapping system to guide radiofrequency ablation of typical atrial flutter. METHODS: A multipoint three-dimensional map of the cavo-tricuspid isthmus, septal and lateral atrial wall was performed in 87 patients prior to and following ablation for typical atrial flutter. Evidence of persisting gaps in the line of block was identified by visual inspection of the color-coded activation maps and these sites were re-ablated. The conduction sequence was also assessed with conventional bidirectional pacing and recording. The assess the reduction in fluoroscopy time, two groups of patients were compared: group A (14 patients) in whom the entire mapping-ablation procedure was guided by the three-dimensional system (Carto, Biosense-Webster, Diamond Bar, CA, USA) and group B (32 patients) in whom the same protocol was used but the procedure was guided by standard fluoroscopic imaging. RESULTS: Acute success was achieved in every patient. During bilateral isthmus pacing, the mean local activation time increased from 20.3 +/- 13.3 ms pre-ablation to 148.3 +/- 53.2 ms (p < 0.01) post-ablation with a mean difference of 120 +/- 31 ms. In 11 patients (9.2%) there was evidence of persisting conduction across the line of block despite evidence of reverse activation of the cavo-tricuspid isthmus by conventional pacing. A gap in the ablation line was identified and re-ablated. At a mean follow-up of 16.3 +/- 2.2 months, there were 5 (4.2%) recurrences of atrial flutter and 12 (10%) recurrences of isolated atrial fibrillation. Four of the 5 recurrences occurred in patients in whom ablation was guided by conventional fluoroscopy (group B). The fluoroscopy time was 4.2 +/- 1.5 min in group A and 27.2 +/- 8.2 min in group B (p < 0.001). CONCLUSIONS: Multipoint mapping of the ablation line following radiofrequency ablation of typical atrial flutter performed using the Carto system allows a more accurate assessment of the isthmus conduction block. This has the potential to reduce the recurrence rate to the level observed for other supraventricular tachycardias.

Atrial Flutter↗

Brain and sense organ anatomy and histology in hemoglobinless Antarctic icefishes (Perciformes: Notothenioidei: Channichthyidae).

The Channichthyidae, one of five Antarctic notothenioid families, includes 16 species and 11 genera. Most live at depths of 200-800 m and are a major component of fish biomass in many shelf areas. Channichthyids are unique among adult fishes in possessing pale white blood containing a few vestigal erythrocytes and no hemoglobin. Here we describe the brains of seven species and special sense organs of eight species of channichthyids. We emphasize Chionodraco hamatus and C. myersi, compare these species to other channichthyids, and relate our findings to what is known about brains and sense organs of red-blooded notothenioids living sympatrically on the Antarctic shelf. Brains of channichthyids generally resemble those of their bathydraconid sister group. Among channichthyids the telencephalon is slightly regressed, resulting in a stalked appearance, but the tectum, corpus cerebellum, and mechanoreceptive areas are well developed. Interspecific variation is present but slight. The most interesting features of channichthyid brains are not in the nervous tissue but in support structures: the vasculature and the subependymal expansions show considerable elaboration. Channichthyids have large accessory nasal sacs and olfactory lamellae are more numerous than in other notothenioids. The eyes are relatively large and laterally oriented with similar duplex (cone and rod) retinae in all eight species. Twin cones are the qualitatively dominant photoreceptor in histological sections and, unlike bathydraconids, there are no species with rod-dominated retinae. Eyes possess the most extensive system of hyaloid arteries known in teleosts. Unlike the radial pattern seen in red-blooded notothenioids and most other teleosts, channichthyid hyaloid arteries arise from four or five main branches and form a closely spaced anastomosing series of parallel channels. Cephalic lateral line canals are membranous and some exhibit extensions (canaliculi), but canals are more ossified than those of deeper-living bathydraconids. We conclude that, with respect to the anatomy and histology of the neural structures, the brain and sensory systems show little that is remarkable compared to other fishes, and exhibit little diversification within the family. Thus, the unusual habitat and a potentially deleterious mutation resulting in a hemoglobinless phenotype are reflected primarily in expansion of the vasculature in the brain and eye partially compensating for the absence of respiratory pigments. Neural morphology gives the impression that channichthyids are a homogeneous and little diversified group.

Adaptation, Physiological↗

Ambulatory computer system for recording and controlling locomotion.

Description of an original miniature computer system capable of recording biomechanical parameters during locomotion, providing auditory signals which can be used in biofeedback locomotor learning, and triggering functional electrical stimulation of nerves and muscles at preselected moments in the locomotor cycle. The system, which is worn on the patient's back, is connected to appropriate gauges with which it is possible to measure knee angle and to record the time of heel contact with the ground. The relevant data are stored in the system memory and can be transferred to a microcassette and later displayed as a function of time on a video screen or printed on paper. The same data can be used on-line to provide a patient with a sound signal as a basis for biofeedback training. This kind of training has been successfully used to correct knee hyperextension during locomotion in hemiplegic patients. The system can also be used to trigger a stimulator and thus constitutes a valuable tool for investigations and treatments involving functional electrical stimulation. Many other applications may be developed, both in basic research and in various fields where motor training is required.

Biofeedback, Psychology↗